EP1229127A1 - Method for the enzymatic preparation of enantiomerically pure derivatives of 1,3-Dioxolan-4-one and 1,3-Oxathiolan-5-one - Google Patents

Method for the enzymatic preparation of enantiomerically pure derivatives of 1,3-Dioxolan-4-one and 1,3-Oxathiolan-5-one Download PDF

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EP1229127A1
EP1229127A1 EP02001124A EP02001124A EP1229127A1 EP 1229127 A1 EP1229127 A1 EP 1229127A1 EP 02001124 A EP02001124 A EP 02001124A EP 02001124 A EP02001124 A EP 02001124A EP 1229127 A1 EP1229127 A1 EP 1229127A1
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alkenyl
alkyl
aryl
heteroaryl
unsubstituted
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EP1229127B1 (en
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Alfred Dr. Popp
Jürgen Dr. Stohrer
Hermann Dr. Petersen
Andrea Gilch
Jodoca Rockinger-Mechlem
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Wacker Chemie AG
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Consortium fuer Elektrochemische Industrie GmbH
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P41/00—Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture
    • C12P41/001—Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture by metabolizing one of the enantiomers

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  • the invention relates to a method for enzymatic production of enantiomerically pure 1,3-dioxolan-4-one and 1,3-oxathiolan-5-one derivatives.
  • Enantiomerically pure derivatives serve as starting materials or Intermediates in the synthesis of agrochemicals and Pharmaceuticals. Many of these connections are currently considered Racemate or mixture of diastereomers produced and marketed. In many cases, the desired physiological effect but only caused by one enantiomer / diastereomer. The other In the best case, isomer is inactive, but it can also counteract the desired effect or even be toxic. Processes for the separation of racemates are therefore always important for the preparation of highly enantiomerically pure compounds.
  • the object of the present invention is to provide a method for Production of enantiomerically pure 1,3-dioxolan-4-one and 1,3-oxathiolan-5-one derivatives to provide which one is inexpensive and avoids the disadvantages mentioned.
  • the object is achieved by a method in which a mixture containing enantiomeric 1,3-dioxolan-4-one or 1,3-oxathiolan-5-one derivatives and a hydrolytically active enzyme are contacted in the presence of a nucleophile, wherein the dioxolanone or oxathiolanone ring of an enantiomer is split by the hydrolytically active enzyme and after cleavage of one enantiomer is the undissolved Enantiomer of the 1,3-dioxolan-4-one or 1,3-oxathiolan-5-one derivative is isolated.
  • the process according to the invention separates a mixture of enantiomers on the dioxolanone or oxathiolanone stage and thus adjusts enantiomerically pure derivative available, which is now in an known the production of an enantiomerically pure 1,3-Dioxolanyl or 1,3-oxathiolanyl nucleoside enables.
  • 1,3-Dioxolan-4-one or 1,3-oxathiolan-5-one have in the ring a hydrolytically labile ester bond, which is catalyzed by an enzyme Reaction can be split. It was surprising found that this ester bond is in the dioxolanone or Oxathiolanone ring with both high enantioselectivity and also high regioselectivity towards others in the connection existing hydrolytically labile groups by a hydrolytically active enzyme cleaves.
  • the method according to the invention is therefore preferred characterized in that a mixture containing enantiomeric 1,3-dioxolan-4-one or 1,3-oxathiolan-5-one derivatives with an enzyme, that is capable of cleaving an ester bond in the presence of a nucleophile of the general formula NuH in contact brings, so that preferably an enantiomer is cleaved.
  • Equation 5 oxygen or sulfur and the radicals R 1 and R 2 are different and are independently selected from the group H, substituted or unsubstituted C 6 -C 18 aryl, C 3 -C 18 heteroaryl, C 1 -C 18 alkyl, C 2 -C 18 alkenyl, C 2 -C 18 alkynyl, C 6 -C 18 aryl-C 1 -C 18 alkyl, C 3 -C 18 heteroaryl-C 1 -C 18 alkyl, C 6 -C 18 - Aryl-C 2 -C 18 alkenyl, C 3 -C 18 heteroaryl-C 2 -C 18 alkenyl, C 1 -C 18 alkoxy-C 1 -C 18 alkyl, C 1 -C 18 alkoxy- C 2 -C 18 alkenyl, C 6 -C 18 aryloxy-C 1 -C 18 alkyl, C 6 -C 18 aryloxy-C 1 -C 18 alkyl,
  • residues are substituted residues, these are preferably alkyl, alkenyl, alkynyl, Aryl, heteroaryl, hydroxy, alkoxy, carboxylate, alkoxycarbonyl, Amino, nitro or halogen radicals substituted.
  • radicals contain a heteroatom, it is preferably O, N or S.
  • Enantiomer mixtures of the general formula (I) are preferably used, wherein R 3 , R 4 , R 8 and R 9 have the meaning already mentioned and R 11 substituted or unsubstituted, branched or unbranched C 1 -C 18 alkyl, C 2 -C 18 alkenyl or C 2 -C 18 alkynyl , substituted or unsubstituted aryl, substituted or unsubstituted silaalkyl or silaaryl or R 11 means COR 10 , where R 10 has the meaning already mentioned.
  • Enantiomer mixtures of the general formula (II) are used with particular preference, wherein R 3 and R 4 have the meaning already mentioned and R 10 is selected from the group substituted or unsubstituted aryl, substituted or unsubstituted, C 1 -C 18 alkyl, C 2 -C 18 alkenyl or C 2 -C 18 - alkynyl.
  • the nucleophile NuH is preferably an oxygen-containing nucleophile OR 5 .
  • all enzymes are for the process according to the invention, which are capable of cleaving an ester bond. It is preferably a lipase or esterase Class 3.1 according to the International Enzyme Nomenclature, Committee of the International Union of Biochemistry and Molecular Biology. Special because of their easier accessibility it is preferably lipases or esterases microbial Origin, swine pancreatic lipase, horse liver esterase or pork liver esterase.
  • Enzymes from fungi, yeasts or bacteria such as Alcaligenes sp., Aspergillus niger, Aspergillus oryzae, Bacillus sp., Bacillus stearothermophilus, Bacillus thermoglucosidasius, Candida antarctica, Candida lipolytica, Candida rugosa, Chromobacterium viscosum, Geotrichium candium, Mucor miehei, Penicillium camembertii, Penicillium roquefortii, Pseudomonas cepacia, Pseudomonas fluorescens, Pseudomonas sp., Rhizomucor javanicus, Rhizopus arrhizus, Rhizopus niveus, Saccharomyces cerevisae, Thermoanaerobium brockii, Thermomyces lanuginosa.
  • Novozym® 435, 525 are very particularly preferred as the enzyme (commercially available from Novo, Denmark), Chirazyme® L2, E1, E2 and L7 (commercially available from Bschreibinger Mannheim, Germany).
  • the enzyme is bound directly in the reaction or as an immobilizate used on various carriers.
  • An immobilized product can be produced in a manner known per se become. This is possible, for example, by dissolving the enzyme in a buffer at a suitable pH and then passively Adsorption on the carrier such.
  • the enzymes can also be covalently attached to the Carrier are bound (e.g. polystyrene or epoxy resins such as Eupergit®). For example, one bound to a carrier in this way Enzyme can be dried by lyophilization.
  • the amount of enzyme to be used in the process according to the invention depends on the type of starting material, product and activity the enzyme preparation.
  • the optimal amount of enzyme for the reaction can be determined by simple preliminary tests.
  • the enzyme-substrate ratio is calculated as Molar ratio between enzyme and dioxolanone / oxathiolanone derivative usually between 1: 1000 and 1: 50000000 or more, preferably at 1: 10000 to 1: 5000000.
  • the method according to the invention can be used both in pure nucleophile (NuH) as a solvent as well as in mixtures of the nucleophile (NuH) with aprotic or protogenic solvents or solvent mixtures are carried out, if these do not affect the reactivity of the hydrolytically active enzyme or lead to undesirable side reactions.
  • Suitable solvents are, for example, aliphatic or aromatic hydrocarbons such as hexane, cyclohexane, petroleum ether or toluene, halogenated hydrocarbons such as methylene chloride or chloroform, ethers such as methyl tert-butyl ether (MTBE), diethyl ether, diisopropyl ether, THF or dioxane, ester, acetonitrile or optionally alcohols that are not a nucleophile in the sense of the above-mentioned enzymatic reaction, such as. B. tertiary alcohols, or mixtures of the compounds mentioned.
  • aliphatic or aromatic hydrocarbons such as hexane, cyclohexane, petroleum ether or toluene
  • halogenated hydrocarbons such as methylene chloride or chloroform
  • ethers such as methyl tert-butyl ether (MTBE), diethyl ether,
  • the nucleophile / solvent ratio (v / v) lies here preferably in a range from 1: 10000 to 1000: 1.
  • nucleophile with aprotic are particularly preferred Solvents such as MTBE or diisopropyl ether in one Nucleophile / solvent ratio (v / v) from 1: 100 to 100 : 1.
  • Solvents such as MTBE or diisopropyl ether in one Nucleophile / solvent ratio (v / v) from 1: 100 to 100 : 1.
  • nucleophile OH
  • this can be adjusted by adding a buffer in order to maintain a predetermined pH.
  • An Na 2 HPO 4 / NaH 2 PO 4 buffer with a pH of 7.0 is preferably used for this.
  • an aqueous alkali preferably the solution of an alkali metal hydroxide in water, particularly preferably the aqueous solution of NaOH or KOH, can be metered in.
  • the reaction is advantageously carried out at a temperature between 0 ° C and 75 ° C, preferably between 10 ° C. and 60 ° C, particularly preferably between 20 ° C and 50 ° C.
  • reaction times are Dioxolanons, choice of nucleophile and solvent and type of enzyme and amount between 10 minutes and 7 days.
  • response times are between 1 and 48 hours.
  • the course of the reaction can be easily carried out using conventional methods track for example by HPLC.
  • the determination can preferably the course of the reaction by measuring the change in optical rotation of the reaction solution in a polarimeter respectively.
  • the course of the reaction is particularly preferably determined online, by measuring the optical rotation value in the a secondary circuit of the reactor.
  • the reaction may vary desired result (high conversion, high enantiomeric excess of the substrate). Ideally, the reaction is with a conversion of 50% with a high enantiomeric purity finished in the substrate.
  • the reaction is preferably carried out, for example, by separation the substrate or the product of the enzyme, e.g. B. by extraction the aqueous phase or filtration ended.
  • the demolition the reaction can also be carried out by deactivating the enzyme, e.g. B. by thermal or chemical denaturation.
  • reaction occurs by repeated, continuous pumping the reaction solution through a container filled with enzyme is carried out (a particularly preferred procedure), the reaction is preferably stopped by pumping around completed.
  • the isolation of the undissolved pure enantiomer takes place preferably by separating off those formed during the reaction By-products and the solvent.
  • the free carbonyl compound R 1 COR 2 formed during the cleavage of a 1,3-dioxolan-4-one or 1,3-oxathiolan-5-one ring and the acid derivative HXCR 3 R 4 CONu can be separated from the reaction solution by simple physical operations , This is preferably done by distillation.
  • the carbonyl compound formed in the enzymatic reaction is an important, expensive precursor in the synthesis of racemic 1,3-dioxolan-4-one or 1,3-oxathiolan-5-one compounds. In order to save chemicals and costs, it is preferably used in the synthesis of 1,3-dioxolan-4-ones and 1,3-oxathiolan-5-ones (see Scheme 1).
  • a polarimeter is connected to the 4-neck flask via a bypass system, with the aid of which the course of the reaction is tracked by measuring the optical rotation value of the solution.
  • reaction monitoring by chiral GC the reaction mixture is filtered off from the undissolved enzyme to terminate the reaction.
  • the reaction mixture is then concentrated in vacuo.
  • the residue is then taken up in 100 mL MTBE and washed twice with 100 mL water.
  • the organic phase is dried over Na 2 SO 4 and then i. Vak. freed from the solvent.

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Abstract

Preparation of enantiomerically pure 1,3-dioxolan-4-one or 1,3-oxathiolan-5-one derivatives (I) involves (i) contacting a mixture of the enantiomers of (I) with a hydrolytic enzyme and a nucleophile, so that the ring of one enantiomer of (I) is cleaved by the enzyme; and (ii) isolating the non-cleaved enantiomer of (I).

Description

Die Erfindung betrifft ein Verfahren zur enzymatischen Herstellung von enantiomerenreinen 1,3-Dioxolan-4-on- und 1,3-Oxathiolan-5-on-Derivaten.The invention relates to a method for enzymatic production of enantiomerically pure 1,3-dioxolan-4-one and 1,3-oxathiolan-5-one derivatives.

Enantiomerenreine Derivate dienen als Ausgangsmaterialien bzw. Zwischenprodukte bei der Synthese von Agrochemikalien und Pharmazeutika. Viele dieser Verbindungen werden zur Zeit als Racemat oder Diastereomerengemisch hergestellt und vermarktet. In vielen Fällen wird der gewünschte physiologische Effekt aber nur von einem Enantiomer/ Diastereomer bewirkt. Das andere Isomer ist im günstigsten Fall inaktiv, es kann aber auch dem gewünschten Effekt entgegenwirken oder sogar toxisch sein. Verfahren zur Trennung von Racematen werden deshalb immer wichtiger für die Darstellung hochenantiomerenreiner Verbindungen.Enantiomerically pure derivatives serve as starting materials or Intermediates in the synthesis of agrochemicals and Pharmaceuticals. Many of these connections are currently considered Racemate or mixture of diastereomers produced and marketed. In many cases, the desired physiological effect but only caused by one enantiomer / diastereomer. The other In the best case, isomer is inactive, but it can also counteract the desired effect or even be toxic. Processes for the separation of racemates are therefore always important for the preparation of highly enantiomerically pure compounds.

Es ist bekannt, dass die Racemattrennung chiraler Verbindungen mit Hilfe von Enzymen durchgeführt werden kann. In einer Vielzahl von Publikationen werden enzymatisch kinetische Racematspaltungen von Estern mit Lipasen und Esterasen beschrieben. Es gibt jedoch bisher kein Verfahren, dass die einfache Trennung von 1,3-Dioxolan-4-on- bzw. 1,3-Oxathiolan-5-on-Derivaten erlaubt. Enantiomerenreine 1,3-Dioxolan-4-one sind von großem Interesse für die Herstellung von anti-viral wirksamen Verbindungen, wie z. B. das 1,3-Dioxolanyl-Nucleosid "Dioxolane-T" (NB = Thymin in Gleichung 1) und ähnlichen Strukturen (Bioorg. Med. Chem. Lett. 1993, 3(2), S. 169-174).

Figure 00010001
It is known that the resolution of chiral compounds can be carried out with the aid of enzymes. A large number of publications describe enzymatic kinetic resolution of esters with lipases and esterases. So far, however, there is no method that allows simple separation of 1,3-dioxolan-4-one or 1,3-oxathiolan-5-one derivatives. Enantiomerically pure 1,3-dioxolan-4-ones are of great interest for the production of anti-viral compounds, such as. B. the 1,3-dioxolanyl nucleoside "Dioxolane-T" (NB = thymine in equation 1) and similar structures ( Bioorg. Med. Chem. Lett. 1993, 3 (2), pp. 169-174).
Figure 00010001

Zur Herstellung von enantiomerenreinen 1,3-Dioxolanyl-Nucleosiden wird die Trennung in die Enantiomere bisher auf der erheblich teureren Nucleosidstufe durchgeführt. Erstmals beschrieben wird dieses Verfahren von L. J. Wilson et al. (Bioorg. Med. Chem. Lett. 1993, 3(2), S. 169-174). Der Buttersäureester der primären Hydroxylgruppe eines 1,3-Dioxolanyl-Nucleosids wird dort mit Hilfe von Schweineleberesterase hydrolysiert und so die beiden reinen Enantiomere in guten optischen Ausbeuten erhalten. WO 00/22157 (Erfinder: Yao, Y. et al.) beschreibt eine Variante dieses Verfahrens durch Resolution in nicht-homogenen Systemen. Enantiomerenreine 1,3-Oxathiolan-5-one sind ebenso von großem Interesse für die Herstellung von anti-viral wirksamen Verbindungen, wie z. B. das 1,3-Oxathiolanyl-Nucleosid Coviracil® (auch Emtricitabine, früher FTC, 4-Amino-5-fluoro-1-[(2R,5S)-2-(hydroxymethyl)-1,3-oxathiolan-5-yl]-2(1H)-pyrimidinone; Gleichung 2) und ähnlichen Strukturen (J. Org. Chem. 1992, 57(21), S. 5563-5565, WO 91/11186, WO 92/14743, WO 00/22157).

Figure 00020001
For the production of enantiomerically pure 1,3-dioxolanyl nucleosides, the separation into the enantiomers has hitherto been carried out at the considerably more expensive nucleoside stage. This method is described for the first time by LJ Wilson et al. ( Bioorg. Med. Chem. Lett. 1993, 3 (2), pp. 169-174). The butyric acid ester of the primary hydroxyl group of a 1,3-dioxolanyl nucleoside is hydrolyzed there with the help of pig liver esterase and the two pure enantiomers are thus obtained in good optical yields. WO 00/22157 (inventor: Yao, Y. et al.) Describes a variant of this method by resolution in non-homogeneous systems. Enantiomerically pure 1,3-oxathiolan-5-ones are also of great interest for the production of anti-viral compounds, such as. B. the 1,3-oxathiolanyl nucleoside Coviracil® (also emtricitabine, formerly FTC, 4-amino-5-fluoro-1 - [(2R, 5S) -2- (hydroxymethyl) -1,3-oxathiolan-5- yl] -2 (1H) -pyrimidinones; equation 2) and similar structures ( J. Org. Chem. 1992, 57 (21), pp. 5563-5565, WO 91/11186, WO 92/14743, WO 00/22157 ).
Figure 00020001

Zur Herstellung von enantiomerenreinen 1,3-Oxathiolanyl-Nucleosiden kann die Trennung in die Enantiomere auf verschiedenen Stufen erfolgen. So ist eine enzymatische Racematspaltung auf der Oxathiolanonstufe möglich. Diese wird von Liotta et al. (WO 91/11186) beschrieben. Dabei wird die Stereoselektion durch eine enzymatische Esterspaltung eines Substituenten in 2-Position des Oxathiolanrings erreicht (Gleichung 3).

Figure 00030001
Als Beispiel wird die Hydrolyse des Buttersäureesters (R = C3H7) in Gegenwart von Schweineleberesterase (PLE) genannt.For the production of enantiomerically pure 1,3-oxathiolanyl nucleosides, the separation into the enantiomers can be carried out at different stages. An enzymatic resolution of racemates at the oxathiolanone stage is thus possible. This is from Liotta et al. (WO 91/11186). Stereoselection is achieved by enzymatic ester cleavage of a substituent in the 2-position of the oxathiolane ring (Equation 3).
Figure 00030001
The hydrolysis of the butyric acid ester (R = C 3 H 7 ) in the presence of pig liver esterase (PLE) is mentioned as an example.

Die zweite Möglichkeit besteht in der Racematspaltung auf der erheblich teureren Nucleosidstufe. Beschrieben wird dieses Verfahren von Liotta et al. (J. Org. Chem. 1992, 57(21), S. 5563-5565 und WO 92/14743). Dabei werden verschiedene Esteracylgruppen des Nucleosidracemats in Gegenwart von Lipasen bzw. Proteasen stereoselektiv abgespalten (Gleichung 4).

Figure 00030002
The second possibility is the resolution of racemates at the considerably more expensive nucleoside level. This method is described by Liotta et al. ( J. Org. Chem. 1992, 57 (21), pp. 5563-5565 and WO 92/14743). Various ester acyl groups of the nucleoside racemate are cleaved stereoselectively in the presence of lipases or proteases (Equation 4).
Figure 00030002

Dabei werden z. T. hohe Enantiomerenüberschüsse (ee(Ester) > 98%)bei guten Ausbeuten (y(Ester) ∼ 45%) erreicht. Eine Verbesserung des Verfahrens aus WO 92/14743 findet sich in WO 00/22157 (Erfinder: Yao, Y. et al.) durch die Verwendung nicht-homogener Reaktionssysteme (Zusatz von nicht-wassermischbaren Cosolventien) zur Racematspaltung von Oxathiolanyl-Nucleosiden.Here, for. T. high enantiomeric excesses (ee (ester)> 98%) with good yields (y (ester) ∼ 45%). An improvement of the method from WO 92/14743 can be found in WO 00/22157 (inventor: Yao, Y. et al.) through the use non-homogeneous reaction systems (addition of non-water-miscible Cosolvents) for resolving oxathiolanyl nucleosides.

Diese Verfahren, die Racematspaltung auf einer sehr späten Stufe durchzuführen, bergen den gravierenden Nachteil eines unnötigen Materialverbrauchs und hohen Anlagenbelegungszeiten, da die maximale Ausbeute einer Racematspaltung bei 50% liegt. This process, the resolution of racemates on a very late Performing a stage have the serious disadvantage of one unnecessary material consumption and high system occupancy times, since the maximum yield of a racemate resolution is 50%.

Die restlichen 50% (die Verbindung mit der falschen Händigkeit) werden im Regelfall verworfen.The remaining 50% (the connection with wrong handedness) are usually rejected.

Aufgabe der vorliegenden Erfindung ist es, ein Verfahren zur Herstellung von enantiomerenreinen 1,3-Dioxolan-4-on- und 1,3-Oxathiolan-5-on-Derivaten zur Verfügung zu stellen, welches kostengünstig ist und die genannten Nachteile vermeidet.The object of the present invention is to provide a method for Production of enantiomerically pure 1,3-dioxolan-4-one and 1,3-oxathiolan-5-one derivatives to provide which one is inexpensive and avoids the disadvantages mentioned.

Die Aufgabe wird gelöst durch ein Verfahren, bei dem ein Gemisch enthaltend enantiomere 1,3-Dioxolan-4-on- bzw. 1,3-Oxathiolan-5-on-Derivate und ein hydrolytisch wirksames Enzym in Gegenwart eines Nucleophils in Kontakt gebracht werden, wobei der Dioxolanon- bzw. Oxathiolanonring eines Enantiomers durch das hydrolytisch wirksame Enzym gespalten wird und nach erfolgter Spaltung des einen Enantiomers das nichtgespaltene Enantiomer des 1,3-Dioxolan-4-on- bzw. 1,3-Oxathiolan-5-on-Derivats isoliert wird.The object is achieved by a method in which a mixture containing enantiomeric 1,3-dioxolan-4-one or 1,3-oxathiolan-5-one derivatives and a hydrolytically active enzyme are contacted in the presence of a nucleophile, wherein the dioxolanone or oxathiolanone ring of an enantiomer is split by the hydrolytically active enzyme and after cleavage of one enantiomer is the undissolved Enantiomer of the 1,3-dioxolan-4-one or 1,3-oxathiolan-5-one derivative is isolated.

Das erfindungsgemäße Verfahren trennt ein Enantiomerengemisch auf der Dioxolanon- bzw. Oxathiolanonstufe und stellt so ein enantiomerenreines Derivat zu Verfügung, welches nun in an sich bekannter Weise die Herstellung eines enantiomerenreinen 1,3-Dioxolanyl- oder 1,3-Oxathiolanyl-Nucleosids ermöglicht.The process according to the invention separates a mixture of enantiomers on the dioxolanone or oxathiolanone stage and thus adjusts enantiomerically pure derivative available, which is now in an known the production of an enantiomerically pure 1,3-Dioxolanyl or 1,3-oxathiolanyl nucleoside enables.

1,3-Dioxolan-4-one bzw. 1,3-Oxathiolan-5-one besitzen im Ring eine hydrolytisch labile Esterbindung, die durch eine enzymkatalysierte Reaktion gespalten werden kann. Überraschend wurde gefunden, dass sich diese Esterbindung im Dioxolanon- bzw. Oxathiolanonring sowohl mit hoher Enantioselektivität, als auch hoher Regioselektivität gegenüber anderen in der Verbindung vorhandenen hydrolytisch labilen Gruppen durch ein hydrolytisch wirksames Enzym spalten lässt.1,3-Dioxolan-4-one or 1,3-oxathiolan-5-one have in the ring a hydrolytically labile ester bond, which is catalyzed by an enzyme Reaction can be split. It was surprising found that this ester bond is in the dioxolanone or Oxathiolanone ring with both high enantioselectivity and also high regioselectivity towards others in the connection existing hydrolytically labile groups by a hydrolytically active enzyme cleaves.

Vorzugsweise ist das erfindungsgemäße Verfahren daher dadurch gekennzeichnet, dass ein Gemisch enthaltend enantiomere 1,3-Dioxolan-4-on- bzw. 1,3-Oxathiolan-5-on-Derivate mit einem Enzym, das zur Spaltung einer Esterbindung befähigt ist in Gegenwart eines Nucleophils der allgemeinen Formel NuH in Kontakt bringt, so dass bevorzugt ein Enantiomer gespalten wird.The method according to the invention is therefore preferred characterized in that a mixture containing enantiomeric 1,3-dioxolan-4-one or 1,3-oxathiolan-5-one derivatives with an enzyme, that is capable of cleaving an ester bond in the presence of a nucleophile of the general formula NuH in contact brings, so that preferably an enantiomer is cleaved.

Diese Spaltung ist schematisch in Gleichung 5 dargestellt,

Figure 00050001
wobei X = Sauerstoff oder Schwefel und
die Reste R1 und R2 ungleich sind und unabhängig voneinander ausgewählt sind aus der Gruppe H, substituiertes oder unsubstituiertes C6-C18-Aryl, C3-C18-Heteroaryl, C1-C18-Alkyl, C2-C18-Alkenyl, C2-C18-Alkinyl, C6-C18-Aryl-C1-C18-Alkyl, C3-C18-Heteroaryl-C1-C18-Alkyl, C6-C18-Aryl-C2-C18-Alkenyl, C3-C18-Heteroaryl-C2-C18-Alkenyl, C1-C18-Alkoxy-C1-C18-Alkyl, C1-C18-Alkoxy-C2-C18-Alkenyl, C6-C18-Aryloxy-C1-C18-Alkyl, C6-C18-Aryloxy-C2-C18-Alkenyl, C3-C8-Cycloalkyl, C3-C8-Cycloalkyl-C1-C18-Alkyl, C3-C8-Cycloalkyl-C2-C18-Alkenyl, CR8R9-On-(CO)m-R10 und die Reste R3 und R4 unabhängig voneinander ausgewählt sind aus der Gruppe substituiertes oder unsubstituiertes C6-C18-Aryl, C3-C18-Heteroaryl, C1-C18-Alkyl, C2-C18-Alkenyl, C2-C18-Alkinyl, C6-C18-Aryl-C1-C18-Alkyl, C3-C18-Heteroaryl-C1-C18-Alkyl, , C6-C18-Aryl-C2-C18-Alkenyl, C3-C18-Heteroaryl-C2-C18-Alkenyl, C1-C18-Alkoxy-C1-C18-Alkyl, C1-C18-Alkoxy-C2-C18-Alkenyl, C6-C18-Aryloxy-C1-C18-Alkyl, C6-C18-Aryloxy-C2-C18-Alkenyl, C3-C8-Cycloalkyl, C3-C8-Cycloalkyl-C1-C18-Alkyl, C3-C8-Cycloalkyl-C2-C18-Alkenyl oder die Reste R3 und R4 zusammen mit dem Kohlenstoff an den sie gebunden sind, ein unsubstituiertes oder substituiertes oder ein Heteroatom enthaltendes Cycloalkyliden bilden, und Nu OR5, SR5, oder NR6R7 bedeutet wobei der Reste R5 ausgewählt sind aus der Gruppe H, substituiertes oder unsubstituiertes C1-C18-Alkyl, C2-C18-Alkenyl, C2-C18-Alkinyl, C6-C18-Aryl-C1-C18-Alkyl, C3-C18-Heteroaryl-C1-C18-Alkyl, C6-C18-Aryl-C2-C18-Alkenyl, C3-C18-Heteroaryl-C2-C18-Alkenyl und die Reste R6 und R7 unabhängig voneinander ausgewählt sind aus der Gruppe H, substituiertes oder unsubstituiertes C1-C18-Alkyl, C2-C18-Alkenyl, C2-C18-Alkinyl, C6-C18-Aryl, C3-C18-Heteroaryl, C6-C18-Aryl-C1-C18-Alkyl, C3-C18-Heteroaryl-C1-C18-Alkyl, C6-C18-Aryl-C2-C18-Alkenyl, C3-C18-Heteroaryl-C2-C18-Alkenyl und
die Reste R8 und R9 unabhängig voneinander ausgewählt sind aus der Gruppe substituiertes oder unsubstituiertes C6-C18-Aryl, C3-C18-Heteroaryl, C1-C18-Alkyl, C2-C18-Alkenyl, C2-C18-Alkinyl, C6-C18-Aryl-C1-C18-Alkyl, C3-C18-Heteroaryl-C1-C18-Alkyl, , C6-C18-Aryl-C2-C18-Alkenyl, C3-C18-Heteroaryl-C2-C18-Alkenyl, C1-C18-Alkoxy-C1-C18-Alkyl, C1-C18-Alkoxy-C2-C18-Alkenyl, C6-C18-Aryloxy-C1-C18-Alkyl, C6-C18-Aryloxy-C2-C18-Alkenyl, C3-C8-Cycloalkyl, C3-C8-Cycloalkyl-C1-C18-Alkyl, C3-C8-Cycloalkyl-C2-C18-Alkenyl oder
die Reste R8 und R9 zusammen mit dem Kohlenstoff an den sie gebunden sind, ein unsubstituiertes oder substituiertes oder ein Heteroatom enthaltendes Cycloalkyliden bilden, und m und n unabhängig voneinander 0 oder 1 bedeuten, und für den Rest R10 gilt:
wenn m = 0 dann ist Rest R10 ausgewählt aus der Gruppe substituiertes oder unsubstituiertes, C1-C18-Alkyl, C2-C18-Alkenyl oder C2-C18-Alkinyl, substituiertes oder unsubstituiertes C6-C18-Aryl, C3-C18-Heteroaryl, substituiertes oder unsubstituiertes Silaalkyl oder Silaaryl, und
wenn m = 1 dann ist Rest R10 ausgewählt aus der Gruppe substituiertes oder unsubstituiertes Aryl, substituiertes oder unsubstituiertes, C1-C18-Alkyl, C2-C18-Alkenyl oder C2-C18-Alkinyl.This split is shown schematically in Equation 5,
Figure 00050001
where X = oxygen or sulfur and
the radicals R 1 and R 2 are different and are independently selected from the group H, substituted or unsubstituted C 6 -C 18 aryl, C 3 -C 18 heteroaryl, C 1 -C 18 alkyl, C 2 -C 18 alkenyl, C 2 -C 18 alkynyl, C 6 -C 18 aryl-C 1 -C 18 alkyl, C 3 -C 18 heteroaryl-C 1 -C 18 alkyl, C 6 -C 18 - Aryl-C 2 -C 18 alkenyl, C 3 -C 18 heteroaryl-C 2 -C 18 alkenyl, C 1 -C 18 alkoxy-C 1 -C 18 alkyl, C 1 -C 18 alkoxy- C 2 -C 18 alkenyl, C 6 -C 18 aryloxy-C 1 -C 18 alkyl, C 6 -C 18 aryloxy-C 2 -C 18 alkenyl, C 3 -C 8 cycloalkyl, C 3 -C 8 cycloalkyl-C 1 -C 18 alkyl, C 3 -C 8 cycloalkyl-C 2 -C 18 alkenyl, CR 8 R 9 -O n - (CO) m -R 10 and the radicals R 3 and R 4 are independently selected from the group of substituted or unsubstituted C 6 -C 18 aryl, C 3 -C 18 heteroaryl, C 1 -C 18 alkyl, C 2 -C 18 alkenyl, C 2 -C 18 -Alkynyl, C 6 -C 18 -aryl-C 1 -C 18 -alkyl, C 3 -C 18 -heteroaryl-C 1 -C 18 -alkyl,, C 6 -C 18 -aryl-C 2 -C 18 - Alkenyl, C 3 - C 18 heteroaryl-C 2 -C 18 alkenyl, C 1 -C 18 alkoxy-C 1 -C 18 alkyl, C 1 -C 18 alkoxy-C 2 -C 18 alkenyl, C 6 -C 18 Aryloxy-C 1 -C 18 alkyl, C 6 -C 18 aryloxy-C 2 -C 18 alkenyl, C 3 -C 8 cycloalkyl, C 3 -C 8 cycloalkyl-C 1 -C 18 alkyl C 3 -C 8 cycloalkyl-C 2 -C 18 alkenyl or the radicals R 3 and R 4 together with the carbon to which they are attached form an unsubstituted or substituted or a heteroatom-containing cycloalkylidene, and Nu OR 5 , SR 5 or NR 6 R 7 means that the radicals R 5 are selected from the group H, substituted or unsubstituted C 1 -C 18 alkyl, C 2 -C 18 alkenyl, C 2 -C 18 alkynyl, C 6 -C 18 aryl-C 1 -C 18 alkyl, C 3 -C 18 heteroaryl-C 1 -C 18 alkyl, C 6 -C 18 aryl-C 2 -C 18 alkenyl, C 3 -C 18 heteroaryl-C 2 -C 18 alkenyl and the radicals R 6 and R 7 are independently selected from the group H, substituted or unsubstituted C 1 -C 18 alkyl, C 2 -C 18 alkenyl, C 2 - C 18 alkynyl, C 6 -C 1 8- aryl, C 3 -C 18 heteroaryl, C 6 -C 18 aryl-C 1 -C 18 alkyl, C 3 -C 18 heteroaryl-C 1 -C 18 alkyl, C 6 -C 18 - Aryl-C 2 -C 18 alkenyl, C 3 -C 18 heteroaryl-C 2 -C 18 alkenyl and
the radicals R 8 and R 9 are independently selected from the group substituted or unsubstituted C 6 -C 18 aryl, C 3 -C 18 heteroaryl, C 1 -C 18 alkyl, C 2 -C 18 alkenyl, C 2 -C 18 alkynyl, C 6 -C 18 aryl-C 1 -C 18 alkyl, C 3 -C 18 heteroaryl-C 1 -C 18 alkyl,, C 6 -C 18 aryl-C 2 -C 18 alkenyl, C 3 -C 18 heteroaryl-C 2 -C 18 alkenyl, C 1 -C 18 alkoxy-C 1 -C 18 alkyl, C 1 -C 18 alkoxy-C 2 -C 18 alkenyl, C 6 -C 18 aryloxy-C 1 -C 18 alkyl, C 6 -C 18 aryloxy-C 2 -C 18 alkenyl, C 3 -C 8 cycloalkyl, C 3 -C 8 - Cycloalkyl-C 1 -C 18 alkyl, C 3 -C 8 cycloalkyl-C 2 -C 18 alkenyl or
the radicals R 8 and R 9 together with the carbon to which they are attached form an unsubstituted or substituted or a heteroatom-containing cycloalkylidene, and m and n independently of one another represent 0 or 1, and the following applies to the radical R 10 :
if m = 0 then radical R 10 is selected from the group substituted or unsubstituted, C 1 -C 18 alkyl, C 2 -C 18 alkenyl or C 2 -C 18 alkynyl, substituted or unsubstituted C 6 -C 18 - Aryl, C 3 -C 18 heteroaryl, substituted or unsubstituted silaalkyl or silaaryl, and
if m = 1 then R 10 is selected from the group substituted or unsubstituted aryl, substituted or unsubstituted, C 1 -C 18 alkyl, C 2 -C 18 alkenyl or C 2 -C 18 alkynyl.

Soweit es sich bei den Resten um substituierte Reste handelt, sind diese vorzugsweise durch Alkyl-, Alkenyl-, Alkinyl-, Aryl-, Heteroaryl-, Hydroxy, Alkoxy-, Carboxylat-, Alkoxycarbonyl-, Amino-, Nitro- oder Halogenreste substituiert.As far as the residues are substituted residues, these are preferably alkyl, alkenyl, alkynyl, Aryl, heteroaryl, hydroxy, alkoxy, carboxylate, alkoxycarbonyl, Amino, nitro or halogen radicals substituted.

Soweit die vorstehend genannten Reste ein Heteroatom enthalten, handelt es sich dabei vorzugsweise um O, N oder S.If the above radicals contain a heteroatom, it is preferably O, N or S.

Bevorzugt Verwendung finden Enantiomerengemische der allgemeinen Formel (I),

Figure 00070001
wobei R3, R4, R8 und R9 die bereits genannte Bedeutung haben und R11 substituiertes oder unsubstituiertes, verzweigtes oder unverzweigtes C1-C18-Alkyl, C2-C18-Alkenyl oder C2-C18-Alkinyl, substituiertes oder unsubstituiertes Aryl, substituiertes oder unsubstituiertes Silaalkyl oder Silaaryl bedeutet oder R11 COR10 bedeutet, wobei R10 die bereits genannte Bedeutung besitzt.Enantiomer mixtures of the general formula (I) are preferably used,
Figure 00070001
wherein R 3 , R 4 , R 8 and R 9 have the meaning already mentioned and R 11 substituted or unsubstituted, branched or unbranched C 1 -C 18 alkyl, C 2 -C 18 alkenyl or C 2 -C 18 alkynyl , substituted or unsubstituted aryl, substituted or unsubstituted silaalkyl or silaaryl or R 11 means COR 10 , where R 10 has the meaning already mentioned.

Besonders bevorzugt Verwendung finden Enantiomerengemische der allgemeinen Formel (II),

Figure 00070002
wobei R3 und R4 die bereits genannte Bedeutung besitzen und R10 ausgewählt ist aus der Gruppe substituiertes oder unsubstituiertes Aryl, substituiertes oder unsubstituiertes, C1-C18-Alkyl, C2-C18-Alkenyl oder C2-C18-Alkinyl.Enantiomer mixtures of the general formula (II) are used with particular preference,
Figure 00070002
wherein R 3 and R 4 have the meaning already mentioned and R 10 is selected from the group substituted or unsubstituted aryl, substituted or unsubstituted, C 1 -C 18 alkyl, C 2 -C 18 alkenyl or C 2 -C 18 - alkynyl.

Das Nucleophil NuH ist bevorzugt ein sauerstoffhaltiges Nucleophil OR5.The nucleophile NuH is preferably an oxygen-containing nucleophile OR 5 .

Besonders bevorzugt handelt es sich bei dem sauerstoffhaltigen Nucleophil um einen niederen, unverzweigten Alkohol, (z. B. Methanol (R5 = CH3) oder Ethanol (R5 = CH2CH3)) oder Wasser (R5 = H).The oxygen-containing nucleophile is particularly preferably a lower, unbranched alcohol (for example methanol (R 5 = CH 3 ) or ethanol (R 5 = CH 2 CH 3 )) or water (R 5 = H).

Für das erfindungsgemäße Verfahren sind prinzipiell alle Enzyme, die zur Spaltung einer Esterbindung befähigt sind, geeignet. Bevorzugt handelt es sich um eine Lipase oder Esterase der Klasse 3.1 gemäß Internationaler Enzym-Nomenklature, Committee of the International Union of Biochemistry and Molecular Biology. Wegen ihrer einfacheren Zugänglichkeit besonders bevorzugt handelt es sich um Lipasen oder Esterasen mikrobiellen Ursprungs, Schweinepankreaslipase, Pferdeleberesterase oder Schweineleberesterase.In principle, all enzymes are for the process according to the invention, which are capable of cleaving an ester bond. It is preferably a lipase or esterase Class 3.1 according to the International Enzyme Nomenclature, Committee of the International Union of Biochemistry and Molecular Biology. Special because of their easier accessibility it is preferably lipases or esterases microbial Origin, swine pancreatic lipase, horse liver esterase or pork liver esterase.

Als Enzyme mikrobiellen Ursprungs seien beispielsweise genannt Enzyme aus Pilzen, Hefen oder Bakterien wie beispielsweise Alcaligenes sp., Aspergillus niger, Aspergillus oryzae, Bacillus sp., Bacillus stearothermophilus, Bacillus thermoglucosidasius, Candida antarctica, Candida lipolytica, Candida rugosa, Chromobacterium viscosum, Geotrichium candium, Mucor miehei, Penicillium camembertii, Penicillium roquefortii, Pseudomonas cepacia, Pseudomonas fluorescens, Pseudomonas sp., Rhizomucor javanicus, Rhizopus arrhizus, Rhizopus niveus, Saccharomyces cerevisae, Thermoanaerobium brockii, Thermomyces lanuginosa. Besonders bevorzugt werden dabei Lipasen und Esterasen aus Candida-Arten wie zum Beispiel Candida antarctica B.Examples of enzymes of microbial origin are mentioned Enzymes from fungi, yeasts or bacteria such as Alcaligenes sp., Aspergillus niger, Aspergillus oryzae, Bacillus sp., Bacillus stearothermophilus, Bacillus thermoglucosidasius, Candida antarctica, Candida lipolytica, Candida rugosa, Chromobacterium viscosum, Geotrichium candium, Mucor miehei, Penicillium camembertii, Penicillium roquefortii, Pseudomonas cepacia, Pseudomonas fluorescens, Pseudomonas sp., Rhizomucor javanicus, Rhizopus arrhizus, Rhizopus niveus, Saccharomyces cerevisae, Thermoanaerobium brockii, Thermomyces lanuginosa. Lipases and esterases are particularly preferred Candida species such as Candida antarctica B.

Als Enzym ganz besonders bevorzugt sind Novozym® 435, 525 (käuflich erhältlich bei Firma Novo, Dänemark), Chirazyme® L2, E1, E2 und L7 (käuflich erhältlich bei Firma Böhringer Mannheim, Deutschland).Novozym® 435, 525 are very particularly preferred as the enzyme (commercially available from Novo, Denmark), Chirazyme® L2, E1, E2 and L7 (commercially available from Böhringer Mannheim, Germany).

Das Enzym wird in der Reaktion direkt oder als Immobilisat gebunden an unterschiedlichste Träger eingesetzt.The enzyme is bound directly in the reaction or as an immobilizate used on various carriers.

Ein Immobilisat kann in an sich bekannter Weise hergestellt werden. Dies ist beispielsweise möglich durch Lösen des Enzyms in einem Puffer bei geeignetem pH und anschließender passiver Adsorption an den Träger wie z. B. Diatomeenerde (Celite®), Aktivkohle, Aluminiumoxid, Kieselgel, Kieselguhr, monodispers lösliche Organosiloxanpartikel oder Harze (z. B. Amberlite®, Dowex®). Alternativ können die Enzyme auch kovalent an den Träger gebunden werden (z. B. Polystyrol oder Epoxy-Harze wie Eupergit®). Ein beispielsweise derart an einen Träger gebundenes Enzym kann durch Lyophilisieren getrocknet werden.An immobilized product can be produced in a manner known per se become. This is possible, for example, by dissolving the enzyme in a buffer at a suitable pH and then passively Adsorption on the carrier such. B. diatomaceous earth (Celite®), Activated carbon, aluminum oxide, silica gel, Kieselguhr, monodisperse soluble organosiloxane particles or resins (e.g. Amberlite®, Dowex®). Alternatively, the enzymes can also be covalently attached to the Carrier are bound (e.g. polystyrene or epoxy resins such as Eupergit®). For example, one bound to a carrier in this way Enzyme can be dried by lyophilization.

Die im erfindungsgemäßen Verfahren einzusetzende Menge an Enzym hängt von der Art des Edukts, Produkts und der Aktivität der Enzympräparation ab. Die für die Reaktion optimale Enzymmenge kann durch einfache Vorversuche ermittelt werden.The amount of enzyme to be used in the process according to the invention depends on the type of starting material, product and activity the enzyme preparation. The optimal amount of enzyme for the reaction can be determined by simple preliminary tests.

Je nach Enzym liegt das Enzym-Substratverhältnis berechnet als Molverhältnis zwischen Enzym und Dioxolanon/Oxathiolanon-Derivat in der Regel zwischen 1 : 1000 und 1 : 50000000 oder mehr, bevorzugt bei 1 : 10000 bis 1 : 5000000.Depending on the enzyme, the enzyme-substrate ratio is calculated as Molar ratio between enzyme and dioxolanone / oxathiolanone derivative usually between 1: 1000 and 1: 50000000 or more, preferably at 1: 10000 to 1: 5000000.

Das erfindungsgemäße Verfahren kann sowohl in reinem Nucleophil (NuH) als Lösungsmittel als auch in Mischungen des Nucleophils (NuH) mit aprotischen oder protogenen Lösungsmitteln oder Lösungsmittelgemischen durchgeführt werden, sofern diese die Reaktivität des hydrolytisch wirksamen Enzyms nicht beeinflussen oder zu unerwünschten Nebenreaktionen führen.The method according to the invention can be used both in pure nucleophile (NuH) as a solvent as well as in mixtures of the nucleophile (NuH) with aprotic or protogenic solvents or solvent mixtures are carried out, if these do not affect the reactivity of the hydrolytically active enzyme or lead to undesirable side reactions.

Vorteilhafterweise wird die Reaktion in einer Mischung aus dem Nucleophil und einem geeigneten Lösungsmittel durchgeführt. Geeignete Lösungsmittel sind beispielsweise aliphatische oder aromatische Kohlenwasserstoffe wie Hexan, Cyclohexan, Petrolether oder Toluol, halogenierte Kohlenwasserstoffe wie Methylenchlorid oder Chloroform, Ether wie Methyl-tert.-Butylether (MTBE), Diethylether, Diisopropylether, THF oder Dioxan, Ester, Acetonitril oder gegebenenfalls Alkohole, die kein Nukleophil im Sinne o. g. enzymatischer Reaktion darstellen, wie z. B. tertiäre Alkohole, oder Mischungen der genannten Verbindungen.The reaction is advantageously carried out in a mixture of the nucleophile and a suitable solvent. Suitable solvents are, for example, aliphatic or aromatic hydrocarbons such as hexane, cyclohexane, petroleum ether or toluene, halogenated hydrocarbons such as methylene chloride or chloroform, ethers such as methyl tert-butyl ether (MTBE), diethyl ether, diisopropyl ether, THF or dioxane, ester, acetonitrile or optionally alcohols that are not a nucleophile in the sense of the above-mentioned enzymatic reaction, such as. B. tertiary alcohols, or mixtures of the compounds mentioned.

Das Verhältnis Nucleophil/ Lösungsmittel (v/v) liegt dabei vorzugsweise in einem Bereich von 1 : 10000 bis 1000 : 1.The nucleophile / solvent ratio (v / v) lies here preferably in a range from 1: 10000 to 1000: 1.

Besonders bevorzugt sind Mischungen des Nucleophils mit aprotische Lösungsmittel, wie MTBE oder Diisopropylether in einem Verhältnis Nucleophil/ Lösungsmittel (v/v) von 1 : 100 bis 100 : 1.Mixtures of the nucleophile with aprotic are particularly preferred Solvents such as MTBE or diisopropyl ether in one Nucleophile / solvent ratio (v / v) from 1: 100 to 100 : 1.

Wird als Nucleophil Wasser verwendet (Nu = OH), so kann, um einen vorgegebenen pH-Wert einzuhalten, dieser durch Zugabe eines Puffers eingestellt werden. Bevorzugt verwendet wird dazu ein Na2HPO4/ NaH2PO4-Puffer mit einem pH von 7,0. Zum selben Zweck kann auch eine wässrige Lauge, bevorzugt die Lösung eines Alkalihydroxyds in Wasser, besonders bevorzugt die wässrige Lösung von NaOH oder KOH, zudosiert werden.If water is used as the nucleophile (Nu = OH), this can be adjusted by adding a buffer in order to maintain a predetermined pH. An Na 2 HPO 4 / NaH 2 PO 4 buffer with a pH of 7.0 is preferably used for this. For the same purpose, an aqueous alkali, preferably the solution of an alkali metal hydroxide in water, particularly preferably the aqueous solution of NaOH or KOH, can be metered in.

Die Reaktion wird vorteilhafterweise bei einer Temperatur zwischen 0 °C und 75 °C durchgeführt, bevorzugt zwischen 10 °C und 60 °C, besonders bevorzugt zwischen 20 °C und 50 °C.The reaction is advantageously carried out at a temperature between 0 ° C and 75 ° C, preferably between 10 ° C. and 60 ° C, particularly preferably between 20 ° C and 50 ° C.

Die Reaktionszeiten betragen je nach Substitutionsmusters des Dioxolanons, Wahl des Nucleophils und Lösungsmittels und Enzymart und -menge zwischen 10 Minuten und 7 Tagen. Bevorzugt liegen die Reaktionszeiten zwischen 1 und 48 Stunden.Depending on the substitution pattern, the reaction times are Dioxolanons, choice of nucleophile and solvent and type of enzyme and amount between 10 minutes and 7 days. Prefers the response times are between 1 and 48 hours.

Der Reaktionsverlauf läßt sich leicht mit üblichen Methoden beispielsweise durch HPLC verfolgen. Bevorzugt kann die Bestimmung des Reaktionsverlaufs durch Messung der Änderung des optischen Drehwerts der Reaktionslösung in einem Polarimeter erfolgen. Besonders bevorzugt erfolgt die Bestimmung des Reaktionsverlaufs online, durch Messung des optischen Drehwerts im einem Nebenkreislauf des Reaktors. Die Reaktion kann je nach gewünschtem Ergebnis (hohe Umsetzung, hoher Enantiomerenüberschuß des Substrats) beendet werden. Im Idealfall ist die Reaktion bei einem Umsatz von 50 % bei einer hohen Enantiomerenreinheit im Substrat beendet.The course of the reaction can be easily carried out using conventional methods track for example by HPLC. The determination can preferably the course of the reaction by measuring the change in optical rotation of the reaction solution in a polarimeter respectively. The course of the reaction is particularly preferably determined online, by measuring the optical rotation value in the a secondary circuit of the reactor. The reaction may vary desired result (high conversion, high enantiomeric excess of the substrate). Ideally, the reaction is with a conversion of 50% with a high enantiomeric purity finished in the substrate.

Vorzugsweise wird die Reaktion beispielsweise durch Separation des Substrats bzw. des Produkts vom Enzym, z. B. durch Extraktion der wässrigen Phase oder Filtration beendet. Der Abbruch der Reaktion kann auch durch Desaktivierung des Enzyms, z. B. durch thermische oder chemische Denaturierung erfolgen.The reaction is preferably carried out, for example, by separation the substrate or the product of the enzyme, e.g. B. by extraction the aqueous phase or filtration ended. The demolition the reaction can also be carried out by deactivating the enzyme, e.g. B. by thermal or chemical denaturation.

Falls die Reaktion durch wiederholtes, kontinuierliches Pumpen der Reaktionslösung durch einen mit Enzym gefüllten Behälter durchgeführt wird, (eine besonders bevorzugte Verfahrensführung), wird die Reaktion vorzugsweise durch Beenden des Umpumpens beendet. If the reaction occurs by repeated, continuous pumping the reaction solution through a container filled with enzyme is carried out (a particularly preferred procedure), the reaction is preferably stopped by pumping around completed.

Die Isolierung des nichtgespaltenen reinen Enantiomers erfolgt vorzugsweise durch Abtrennung der bei der Reaktion entstandenen Nebenprodukte und des Lösungsmittels.The isolation of the undissolved pure enantiomer takes place preferably by separating off those formed during the reaction By-products and the solvent.

Die bei der Spaltung eines 1,3-Dioxolan-4-on- oder 1,3-Oxathiolan-5-onrings entstehende freie Carbonylverbindung R1COR2 und das Säurederivat HXCR3R4CONu können aus der Reaktionslösung durch einfache physikalische Operationen abgetrennt werden. Vorzugsweise geschieht dies durch Destillation.The free carbonyl compound R 1 COR 2 formed during the cleavage of a 1,3-dioxolan-4-one or 1,3-oxathiolan-5-one ring and the acid derivative HXCR 3 R 4 CONu can be separated from the reaction solution by simple physical operations , This is preferably done by distillation.

Darüber hinaus können weitere Abbauprodukte, die bei der Spaltung weiterer funktioneller Gruppen im Molekül gebildet werden leicht abgetrennt werden. Vorzugsweise geschieht dies durch Destillation.In addition, other breakdown products can be used in the cleavage further functional groups are formed in the molecule can be easily separated. This is preferably done by Distillation.

Bevorzugt werden zunächst die niedrigsiedenden Verbindungen destillativ abgetrennt. Überraschend wurde gefunden, dass der Alkohol (R1 = H, R2 = CH2OH), der als Nebenprodukt bei der Racematspaltung eines Esterdioxolanons (X = O; R1 = H, R2 = CH2-O-(CO)-R10) entsteht, durch einfache Extraktion, bevorzugt mit Wasser, abgetrennt werden kann.The low-boiling compounds are preferably first separated off by distillation. Surprisingly, it was found that the alcohol (R 1 = H, R 2 = CH 2 OH), which is a by-product in the resolution of an ester dioxolanone (X = O; R 1 = H, R 2 = CH 2 -O- (CO) -R 10 ) arises, can be separated by simple extraction, preferably with water.

Die bei der enzymatischen Reaktion entstehende Carbonylverbindung ist eine wichtige, teure Vorstufe bei der Synthese von racemischen 1,3-Dioxolan-4-on- bzw. 1,3-Oxathiolan-5-onverbindungen. Sie wird, um Chemikalien und Kosten zu sparen, bevorzugt in die Synthese der 1,3-Dioxolan-4-one bzw. 1,3-Oxathiolan-5-one eingesetzt (s. Schema 1).

Figure 00120001
The carbonyl compound formed in the enzymatic reaction is an important, expensive precursor in the synthesis of racemic 1,3-dioxolan-4-one or 1,3-oxathiolan-5-one compounds. In order to save chemicals and costs, it is preferably used in the synthesis of 1,3-dioxolan-4-ones and 1,3-oxathiolan-5-ones (see Scheme 1).
Figure 00120001

Die folgenden Beispiele dienen der weiteren Beschreibung der Erfindung.The following examples serve to further describe the Invention.

Beispiel 1: (+) - (R)-2-Methylpropansäure (4-Oxo-1,3-dioxolan-2-yl)methylester (Batch-Prozess)Example 1: (+) - ( R ) -2-methylpropanoic acid (4-oxo-1,3-dioxolan-2-yl) methyl ester (batch process)

In einem 1 L 4-Halskolben werden 50.0 g (0.27 mol) racemischer 2-Methylpropansäure (4-Oxo-1,3-dioxolan-2-yl)methylester (X = O; R1 = H; R2 = CH2-O-(CO)-CH(CH3)2; R3, R4 = H) in einer Mischung aus 185 mL MTBE und 185 mL Methanol (Nu = OCH3) gelöst. Zu dieser Lösung gibt man 2.6 g Novozym® 435 und rührt die Mischung heftig.50.0 g (0.27 mol) of racemic 2-methylpropanoic acid (4-oxo-1,3-dioxolan-2-yl) methyl ester (X = O; R 1 = H; R 2 = CH 2 - O- (CO) -CH (CH 3 ) 2 ; R 3 , R 4 = H) dissolved in a mixture of 185 mL MTBE and 185 mL methanol (Nu = OCH 3 ). 2.6 g of Novozym® 435 are added to this solution and the mixture is stirred vigorously.

An den 4-Halskolben ist über ein Bypass-System ein Polarimeter angeschlossen, mit dessen Hilfe der Reaktionsverlauf anhand der Messung des optischen Drehwerts der Lösung verfolgt wird. Bei erreichen der gewünschten Enantiomerenreinheit (Reaktionsverfolgung durch chiral-GC) wird zur Beendigung der Reaktion die Reaktionsmischung vom ungelösten Enzym abfiltriert. Anschließend wird die Reaktionsmischung im Vakuum eingeengt. Der Rückstand wird anschließend in 100 mL MTBE aufgenommen und 2 mal mit je 100 mL Wasser gewaschen. Die organische Phase wird über Na2SO4 getrocknet und dann i. Vak. vom Lösungsmittel befreit. Die Reinigung des Rohprodukts erfolgt durch Destillation.
Ausbeute: 10.3 g (0.05 mol; 20 %)
Sdp.: 55 °C (0.02 mbar) [α]20 D = +19.8 (neat); ee > 98 % A polarimeter is connected to the 4-neck flask via a bypass system, with the aid of which the course of the reaction is tracked by measuring the optical rotation value of the solution. When the desired enantiomeric purity is reached (reaction monitoring by chiral GC), the reaction mixture is filtered off from the undissolved enzyme to terminate the reaction. The reaction mixture is then concentrated in vacuo. The residue is then taken up in 100 mL MTBE and washed twice with 100 mL water. The organic phase is dried over Na 2 SO 4 and then i. Vak. freed from the solvent. The crude product is purified by distillation.
Yield: 10.3 g (0.05 mol; 20%)
Sdp .: 55 ° C (0.02 mbar) [Α] 20 D = +19.8 (neat); ee> 98%

Beispiel 2: (+)-(R)-2-Methylpropansäure (4-Oxo-1,3-dioxolan-2-yl)methylester (Säulen-Prozess)Example 2: (+) - ( R ) -2-methylpropanoic acid (4-oxo-1,3-dioxolan-2-yl) methyl ester (column process)

In einem thermostatisierten 0,6 L Glaskolben werden 50.0 g (0.27 mol) racemischer 2-Methylpropansäure (4-Oxo-1,3-dioxolan-2-yl)methylester (X = O; R1 = H; R2 = CH2-O-(CO)-CH(CH3)2; R3, R4 = H) in einer Mischung aus 185 mL MTBE und 185 mL Methanol (Nu = OCH3) gelöst. In eine separate Glassäule füllt man 1.3 g Novozym® 435 und pumpt über ein Schlauchsystem das Substrat-Lösungsmittelgemisch durch die Glassäule (Flußgeschwindigkeit 600 mL/h).
Zum Stoppen der Reaktion (nach ∼ 25 h) wird das Umpumpen beendet, und das Rohprodukt wird wie in Beispiel 1 beschrieben gereinigt.50.0 g (0.27 mol) of racemic 2-methylpropanoic acid (4-oxo-1,3-dioxolan-2-yl) methyl ester (X = O; R 1 = H; R 2 = CH 2 ) are placed in a thermostated 0.6 L glass flask -O- (CO) -CH (CH 3 ) 2 ; R 3 , R 4 = H) dissolved in a mixture of 185 mL MTBE and 185 mL methanol (Nu = OCH 3 ). 1.3 g of Novozym® 435 is poured into a separate glass column and the substrate-solvent mixture is pumped through the glass column (flow rate 600 mL / h) via a hose system.
To stop the reaction (after ∼ 25 h), the pumping is stopped and the crude product is purified as described in Example 1.

Beispiele 3-8:Examples 3-8:

Folgende Beispiele wurden entsprechend der Vorschrift in Beispiel 1 durchgeführt.

Figure 00140001
The following examples were carried out in accordance with the instructions in Example 1.
Figure 00140001

Claims (14)

Verfahren zur Herstellung eines enantiomerenreinen 1,3-Dioxolan-4-on- bzw. 1,3-Oxathiolan-5-on-Derivats dadurch gekennzeichnet, daß ein Gemisch enthaltend enantiomere 1,3-Dioxolan-4-on- bzw. 1,3-Oxathiolan-5-on-Derivate und ein hydrolytisch wirksames Enzym in Gegenwart eines Nucleophils in Kontakt gebracht werden und der 1,3-Dioxolan-4-on- bzw. 1,3-Oxathiolan-5-on-ring eines Enantiomers durch das hydrolytisch wirksame Enzym gespalten wird und nach erfolgter Spaltung des einen Enantiomers das nicht gespaltene Enantiomer des 1,3-Dioxolan-4-on- bzw. 1,3-Oxathiolan-5-on-Derivats isoliert wird.Process for the preparation of an enantiomerically pure 1,3-dioxolan-4-one or 1,3-oxathiolan-5-one derivative, characterized in that a mixture comprising enantiomeric 1,3-dioxolan-4-one or 1, 3-Oxathiolan-5-one derivatives and a hydrolytically active enzyme in contact in the presence of a nucleophile and the 1,3-dioxolan-4-one or 1,3-oxathiolan-5-one ring of an enantiomer the hydrolytically active enzyme is cleaved and, after cleavage of the one enantiomer, the undissolved enantiomer of the 1,3-dioxolan-4-one or 1,3-oxathiolan-5-one derivative is isolated. Verfahren gemäß Anspruch 1, dadurch gekennzeichnet, dass das Gemisch enthaltend enantiomere 1,3-Dioxolan-4-on bzw. 1,3-Oxathiolan-5-on-Derivate mittels eines Enzyms, das zur Spaltung einer Esterbindung befähigt ist in Gegenwart eines Nucleophils (NuH) wie in Gleichung dargestellt gespalten wird,
Figure 00150001
wobei X = Sauerstoff oder Schwefel und
die Reste R1 und R2 ungleich sind und unabhängig voneinander ausgewählt sind aus der Gruppe H, substituiertes oder unsubstituiertes C6-C18-Aryl, C3-C18-Heteroaryl, C1-C18-Alkyl, C2-C18-Alkenyl, C2-C18-Alkinyl, C6-C18-Aryl-C1-C18-Alkyl, C3-C18-Heteroaryl-C1-C18-Alkyl, C6-C18-Aryl-C2-C18-Alkenyl, C3-C18-Heteroaryl-C2-C18-Alkenyl, C1-C18-Alkoxy-C1-C18-Alkyl, C1-C18-Alkoxy-C2-C18-Alkenyl, C6-C18-Aryloxy-C1-C18-Alkyl, C6-C18-Aryloxy-C2-C18-Alkenyl, C3-C8-Cycloalkyl, C3-C8-Cycloalkyl-C1-C18-Alkyl, C3-C8-Cycloalkyl-C2-C18-Alkenyl, CR8R9-On-(CO)m-R10 und
die Reste R3 und R4 unabhängig voneinander ausgewählt sind aus der Gruppe substituiertes oder unsubstituiertes C6-C18-Aryl, C3-C18-Heteroaryl, C1-C18-Alkyl, C2-C18-Alkenyl, C2-C18-Alkinyl, C6-C18-Aryl-C1-C18-Alkyl, C3-C18-Heteroaryl-C1-C18-Alkyl, , C6-C18-Aryl-C2-C18-Alkenyl, C3-C18-Heteroaryl-C2-C18-Alkenyl, C1-C18-Alkoxy-C1-C18-Alkyl, C1-C18-Alkoxy-C2-C18-Alkenyl, C6-C18-Aryloxy-C1-C18-Alkyl, C6-C18-Aryloxy-C2-C18-Alkenyl, C3-C8-Cycloalkyl, C3-C8-Cycloalkyl-C1-C18-Alkyl, C3-C8-Cycloalkyl-C2-C18-Alkenyl oder die Reste R3 und R4 zusammen mit dem Kohlenstoff an den sie gebunden sind, ein unsubstituiertes oder substituiertes oder ein Heteroatom enthaltendes Cycloalkyliden bilden, und Nu OR5, SR5, oder NR6R7 bedeutet wobei
der Rest R5 ausgewählt ist aus der Gruppe H, substituiertes oder unsubstituiertes C1-C18-Alkyl, C2-C18-Alkenyl, C2-C18-Alkinyl, C6-C18-Aryl-C1-C18-Alkyl, C3-C18-Heteroaryl-C1-C18-Alkyl, C6-C18-Aryl-C2-C18-Alkenyl, C3-C18-Heteroaryl-C2-C18-Alkenyl und
die Reste R6 und R7 unabhängig voneinander ausgewählt sind aus der Gruppe H, substituiertes oder unsubstituiertes C1-C18-Alkyl, C2-C18-Alkenyl, C2-C18-Alkinyl, C6-C18-Aryl, C3-C18-Heteroaryl, C6-C18-Aryl-C1-C18-Alkyl, C3-C18-Heteroaryl-C1-C18-Alkyl, C6-C18-Aryl-C2-C18-Alkenyl, C3-C18-Heteroaryl-C2-C18-Alkenyl und
die Reste R8 und R9 unabhängig voneinander ausgewählt sind aus der Gruppe substituiertes oder unsubstituiertes C6-C18-Aryl, C3-C18-Heteroaryl, C1-C18-Alkyl, C2-C18-Alkenyl, C2-C18-Alkinyl, C6-C18-Aryl-C1-C18-Alkyl, C3-C18-Heteroaryl-C1-C18-Alkyl, C6-C18-Aryl-C2-C18-Alkenyl, C3-C18-Heteroaryl-C2-C18-Alkenyl, C1-C18-Alkoxy-C1-C18-Alkyl, C1-C18-Alkoxy-C2-C18-Alkenyl, C6-C18-Aryloxy-C1-C18-Alkyl, C6-C18-Aryloxy-C2-C18-Alkenyl, C3-C8-Cycloalkyl, C3-C8-Cycloalkyl-C1-C18-Alkyl, C3-C8-Cycloalkyl-C2-C18-Alkenyl oder die Reste R8 und R9 zusammen mit dem Kohlenstoff an den sie gebunden sind, ein unsubstituiertes oder substituiertes oder ein Heteroatom enthaltendes Cycloalkyliden bilden, und
m und n unabhängig voneinander 0 oder 1 bedeuten, und für den Rest R10 gilt:
wenn m = 0 dann ist Rest R10 ausgewählt aus der Gruppe substituiertes oder unsubstituiertes, C1-C18-Alkyl, C2-C18-Alkenyl oder C2-C18-Alkinyl, substituiertes oder unsubstituiertes C6-C18-Aryl, C3-C18-Heteroaryl, substituiertes oder unsubstituiertes Silaalkyl oder Silaaryl, und
wenn m = 1 dann ist Rest R10 ausgewählt aus der Gruppe substituiertes oder unsubstituiertes Aryl, substituiertes oder unsubstituiertes, C1-C18-Alkyl, C2-C18-Alkenyl oder C2-C18-Alkinyl.A method according to claim 1, characterized in that the mixture containing enantiomeric 1,3-dioxolan-4-one or 1,3-oxathiolan-5-one derivatives by means of an enzyme which is capable of breaking an ester bond in the presence of a nucleophile (NuH) is split as shown in the equation,
Figure 00150001
where X = oxygen or sulfur and
the radicals R 1 and R 2 are different and are independently selected from the group H, substituted or unsubstituted C 6 -C 18 aryl, C 3 -C 18 heteroaryl, C 1 -C 18 alkyl, C 2 -C 18 alkenyl, C 2 -C 18 alkynyl, C 6 -C 18 aryl-C 1 -C 18 alkyl, C 3 -C 18 heteroaryl-C 1 -C 18 alkyl, C 6 -C 18 - Aryl-C 2 -C 18 alkenyl, C 3 -C 18 heteroaryl-C 2 -C 18 alkenyl, C 1 -C 18 alkoxy-C 1 -C 18 alkyl, C 1 -C 18 alkoxy- C 2 -C 18 alkenyl, C 6 -C 18 aryloxy-C 1 -C 18 alkyl, C 6 -C 18 aryloxy-C 2 -C 18 alkenyl, C 3 -C 8 cycloalkyl, C 3 -C 8 cycloalkyl-C 1 -C 18 alkyl, C 3 -C 8 cycloalkyl-C 2 -C 18 alkenyl, CR 8 R 9 -O n - (CO) m -R 10 and
the radicals R 3 and R 4 are independently selected from the group of substituted or unsubstituted C 6 -C 18 aryl, C 3 -C 18 heteroaryl, C 1 -C 18 alkyl, C 2 -C 18 alkenyl, C 2 -C 18 alkynyl, C 6 -C 18 aryl-C 1 -C 18 alkyl, C 3 -C 18 heteroaryl-C 1 -C 18 alkyl,, C 6 -C 18 aryl-C 2 -C 18 alkenyl, C 3 -C 18 heteroaryl-C 2 -C 18 alkenyl, C 1 -C 18 alkoxy-C 1 -C 18 alkyl, C 1 -C 18 alkoxy-C 2 -C 18 alkenyl, C 6 -C 18 aryloxy-C 1 -C 18 alkyl, C 6 -C 18 aryloxy-C 2 -C 18 alkenyl, C 3 -C 8 cycloalkyl, C 3 -C 8 - Cycloalkyl-C 1 -C 18 alkyl, C 3 -C 8 cycloalkyl-C 2 -C 18 alkenyl or the radicals R 3 and R 4 together with the carbon to which they are attached, an unsubstituted or substituted or a heteroatom form containing cycloalkylidene, and Nu means OR 5 , SR 5 , or NR 6 R 7 means
the radical R 5 is selected from the group H, substituted or unsubstituted C 1 -C 18 alkyl, C 2 -C 18 alkenyl, C 2 -C 18 alkynyl, C 6 -C 18 aryl-C 1 -C 18 alkyl, C 3 -C 18 heteroaryl-C 1 -C 18 alkyl, C 6 -C 18 aryl-C 2 -C 18 alkenyl, C 3 -C 18 heteroaryl-C 2 -C 18 - Alkenyl and
the radicals R 6 and R 7 are independently selected from the group H, substituted or unsubstituted C 1 -C 18 alkyl, C 2 -C 18 alkenyl, C 2 -C 18 alkynyl, C 6 -C 18 aryl , C 3 -C 18 heteroaryl, C 6 -C 18 aryl-C 1 -C 18 alkyl, C 3 -C 18 heteroaryl-C 1 -C 18 alkyl, C 6 -C 18 aryl-C 2 -C 18 alkenyl, C 3 -C 18 heteroaryl-C 2 -C 18 alkenyl and
the radicals R 8 and R 9 are independently selected from the group substituted or unsubstituted C 6 -C 18 aryl, C 3 -C 18 heteroaryl, C 1 -C 18 alkyl, C 2 -C 18 alkenyl, C 2 -C 18 alkynyl, C 6 -C 18 aryl-C 1 -C 18 alkyl, C 3 -C 18 heteroaryl-C 1 -C 18 alkyl, C 6 -C 18 aryl-C 2 - C 18 alkenyl, C 3 -C 18 heteroaryl-C 2 -C 18 alkenyl, C 1 -C 18 alkoxy-C 1 -C 18 alkyl, C 1 -C 18 alkoxy-C 2 -C 18 -Alkenyl, C 6 -C 18 aryloxy-C 1 -C 18 alkyl, C 6 -C 18 aryloxy-C 2 -C 18 alkenyl, C 3 -C 8 cycloalkyl, C 3 -C 8 cycloalkyl -C 1 -C 18 alkyl, C 3 -C 8 cycloalkyl-C 2 -C 18 alkenyl or the radicals R 8 and R 9 together with the carbon to which they are attached, an unsubstituted or substituted or containing a hetero atom Form cycloalkylidene, and
m and n are independently 0 or 1, and the following applies to the radical R 10 :
if m = 0 then radical R 10 is selected from the group substituted or unsubstituted, C 1 -C 18 alkyl, C 2 -C 18 alkenyl or C 2 -C 18 alkynyl, substituted or unsubstituted C 6 -C 18 - Aryl, C 3 -C 18 heteroaryl, substituted or unsubstituted silaalkyl or silaaryl, and
if m = 1 then R 10 is selected from the group substituted or unsubstituted aryl, substituted or unsubstituted, C 1 -C 18 alkyl, C 2 -C 18 alkenyl or C 2 -C 18 alkynyl. Verfahren gemäß Anspruch 1 oder 2, dadurch gekennzeichnet, dass das hydrolytisch wirksame Enzym eine Lipase oder Esterase ist.A method according to claim 1 or 2, characterized in that the hydrolytically active enzyme is a lipase or esterase. Verfahren gemäß einem der Ansprüche 1 bis 3, dadurch gekennzeichnet, dass das Enzym direkt oder als Immobilisat eingesetzt wird.Method according to one of claims 1 to 3, characterized in that the enzyme is used directly or as an immobilizate. Verfahren gemäß einem der Ansprüche 1 bis 4, dadurch gekennzeichnet, dass Enzym zu Dioxolanon/Oxathiolanon-Derivat berechnet als Molverhältnis zwischen Enzym und Dioxolanon/Oxathiolanon-Derivat in einem Verhältnis von 1 : 1000 bis 1 : 50000000 liegt.Method according to one of claims 1 to 4, characterized in that the enzyme to dioxolanone / oxathiolanone derivative calculated as the molar ratio between the enzyme and dioxolanone / oxathiolanone derivative is in a ratio of 1: 1000 to 1: 50000000. Verfahren gemäß einem der Ansprüche 1 bis 5, dadurch gekennzeichnet, dass das Nucleophil ein sauerstoffhaltiges Nucleophil ist.Method according to one of claims 1 to 5, characterized in that the nucleophile is an oxygen-containing nucleophile. Verfahren gemäß Anspruch 6, dadurch gekennzeichnet, dass das sauerstoffhaltige Nucleophil ein niederer unverzweigter Alkohol oder Wasser ist.A method according to claim 6, characterized in that the oxygen-containing nucleophile is a lower unbranched alcohol or water. Verfahren gemäß Anspruch 7, dadurch gekennzeichnet, dass der niedere unverzweigte Alkohol Methanol oder Ethanol ist.A method according to claim 7, characterized in that the lower unbranched alcohol is methanol or ethanol. Verfahren gemäß einem der Ansprüche 1 bis 8, dadurch gekennzeichnet, dass es in Gegenwart eines Cosolvens durchgeführt wird.Method according to one of claims 1 to 8, characterized in that it is carried out in the presence of a cosolvent. Verfahren gemäß Anspruch 9, dadurch gekennzeichnet, dass das Cosolvens ausgewählt ist aus der Gruppe aliphatische Kohlenwasserstoffe, aromatische Kohlenwasserstoffe, halogenierte Kohlenwasserstoffe, Ether, Alkohole, Ester oder Acetonitril oder Mischungen der genannten Verbindungen.A method according to claim 9, characterized in that the cosolvent is selected from the group aliphatic hydrocarbons, aromatic hydrocarbons, halogenated hydrocarbons, ethers, alcohols, esters or acetonitrile or mixtures of the compounds mentioned. Verfahren gemäß einem der Ansprüche 1 bis 10, dadurch gekennzeichnet, dass die Reaktion bei Temperaturen zwischen 0 und 75 °C durchgeführt wird.Method according to one of claims 1 to 10, characterized in that the reaction is carried out at temperatures between 0 and 75 ° C. Verfahren gemäß einem der Ansprüche 1 bis 11, dadurch gekennzeichnet, dass die Reaktion zwischen 10 Minuten und bis zu 7 Tagen durchgeführt wird.Method according to one of claims 1 to 11, characterized in that the reaction is carried out between 10 minutes and up to 7 days. Verfahren gemäß einem der Ansprüche 1 bis 12, dadurch gekennzeichnet, dass die Isolierung des nichtgespaltenen Enantiomers durch Abtrennung der bei der Reaktion entstandenen Nebenprodukte und des Lösungsmittels erfolgt.Method according to one of claims 1 to 12, characterized in that the isolation of the non-cleaved enantiomer is carried out by separating the by-products formed in the reaction and the solvent. Verfahren nach Anspruch 13 dadurch gekennzeichnet, daß die Nebenprodukte durch Extraktion und Destillation abgetrennt werden.A method according to claim 13, characterized in that the by-products are separated by extraction and distillation.
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